Method for preparing hard-tissue low-temperature buried cold-ester

A technology of hard tissue and low temperature, which is applied in the field of preparation of low-temperature embedding agent for hard tissue, which can solve the problems of not developing low-temperature embedding materials and achieve the effect of easy removal, short time consumption and high hardness

Inactive Publication Date: 2006-08-09
WUHAN UNIV
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  • Summary
  • Abstract
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Problems solved by technology

There is almost no research on low-temperature embedding of hard tissues, let alone the development of corresponding low-temperature embedding materials

Method used

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Examples

Experimental program
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Embodiment Construction

[0024] A kind of hard tissue low-temperature embedding agent——Pyrene Glycerol (PMG) The preparation steps are as follows:

[0025] The technical process of cryogenic embedding of hard tissue is different from that of paraffin embedding.

[0026] 1. Cut the bone tissue into a thickness of 1 to 2 cm with diamond blades. During the sawing process, rinse with cold running water, which requires fast speed. After completion, put it into pre-cooled fixative and store it at 4°C. Fixative configuration and fixation steps are as follows:

[0027]1.6% paraformaldehyde for 24 hours at 4°C—fresh refrigerated fixative (PH7.4) prepared in phosphate buffer prepared with 5% sucrose (recipe: 1 part of 8% paraformaldehyde, 1 part of 0.04M phosphate buffer and 10% sucrose (mixed with pH 7.4, 2 parts of double distilled water) for 24 hours—after the fixation was completed, the tissue was washed overnight at 4°C, pH 7.4, in 0.02M phosphate buffer.

[0028] 2. Alcohol gradient dehydration for wash...

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Abstract

This invention discloses a preparation method for embedding condensing ester at low temperature to hard tissues including: first of all cutting a bone tissue into slices of 1-2cm thick with a diamond-impregnated blade, then putting them into a stationary liquid to be kept, then dehydrating the cleaned sample in gradient with alcohol, then putting the dehydrated sample into an intermediated solution of penetration and then to a penetration solution for vacuumizing in intermission, after that, putting the penetrated tissue into an embedded bottle added with fresh embedding liquid to be polymerized in a refrigerator, then crushing the bottle to take out the block to be cut on a hard tissue slicer to be attached on a glass carrier to be dried and finally dying it .

Description

technical field [0001] The invention relates to a preparation method of a hard tissue low-temperature embedding agent (condensate PMG), which is widely used in research in the fields of medicine, material science, biology and the like. Background technique [0002] The development of hard tissue sections in foreign countries (mainly concentrated in the Nordic countries) was early, and this technology was carried out in many laboratories in the 1980s, and the conventional embedding method of methyl methacrylate (MMA) has been used (30 degrees ~ Polymerization at 40 degrees), methyl methacrylate (MMA) can be removed very easily and completely, and the dyeing result after film production is excellent, and the shape details are good (Erben 1997, Embedding of bone samples inmethylmethacrylate: an improved suitable for bonehistomorphometry, historychemistry , and immunohistochemistry. J Histochem Cytochem). It can be considered that methyl methacrylate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/04
Inventor 陈东张旗
Owner WUHAN UNIV
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